Method for synthesising templated molecules

a templated molecule and synthesis method technology, applied in the direction of nucleotide libraries, electric generator control, machines/engines, etc., can solve the problems of cumbersome split-and-combine methods, difficult generation of molecules carrying new properties, and inability to achieve one-pot amplification of library members, etc., to achieve a higher ratio of complexes

Active Publication Date: 2005-10-06
NUEVOLUTION AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0087] v) obtaining an enriched library having a higher ratio of complexes comprising templated molecules with the predetermined activity.
[0088] The amplification step is normally preferred, though not always necessary. Especially, when several cycles of enrichments are conducted it is of advantage to make an amplification to obtain sufficient complexes. In a preferred aspect of the invention, the amplification of the complexes of the enriched library comp

Problems solved by technology

The generation of molecules carrying new properties remains a challenging task.
This approach does not, however, allow one-pot amplification of the library members.
Furthermore, the sequence

Method used

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  • Method for synthesising templated molecules
  • Method for synthesising templated molecules
  • Method for synthesising templated molecules

Examples

Experimental program
Comparison scheme
Effect test

example 2

[0271] Mix 2 μl Buffer A, 2 μl relevant oligo 1 (0.2 pmol / ul), 1 μl relevant oligo 2 (10 pmol / ul), 1 μl relevant oligo 3 (10 pmol / ul), 4 μl H2O. (See table II, below)

TABLE IIOligo 1 (32P-Experimentlabelled)Oligo 2Oligo 3GAh 5Ah 6NoneHAh 5Ah 6Ah 7IAh 5Ah 6Ah 8JAh 5Ah 6Ah 9KAh 5Ah 6Ah 11

[0272] Anneal as described above. Add 1 μl 100 mM, 10 mM or 1 mM TSAT (Tris-succinimidyl aminotriacetate, Pierce cat#33063 dissolved in DMSO). Incubate at 25° C. for about 5 h, then run 10% urea polyacrylamide gel, as described above.

[0273] The results are shown in FIG. 2

example 3

[0274] Mix 2 μl Buffer A, 2 μl relevant oligo 1 ( 0.2 pmol / ul), 1 μl relevant oligo 2 (10 pmol / ul), 1 μl relevant oligo 3 (10 pmol / ul), 4 μl H2O. (See table III, below)

TABLE IIIOligo 1 (32P-Experimentlabelled)Oligo 2Oligo 3LAh 1Ah 6NoneMAh 1Ah 6Ah 7NAh 1Ah 6Ah 8OAh 1Ah 6Ah 9PAh 1Ah 6Ah 11

[0275] Anneal as described above. Add 1 μl 1M, 100 mM, 10 mM or 1 mM EDC (1-Ethyl-3-(3-dimethylaminopropyl) Carbodiimide Hydrochloride, Fluka #03450) and 1 μl 100 mM NHS (N-Hydroxysuccinimid) (Aldrich cat #13.067-2). Incubation at 25° C. for about 5 h, and analyze by 10% urea polyacrylamide gel electrophoresis, as described above.

[0276] The results are shown in FIG. 3.

example 4

[0277] Mix 2 μl buffer A, B, C, D, E or F, 2 μl relevant oligo 1 ( 0.2 pmol / ul), 1 μl relevant oligo 2 (10 pmol / ul), 1 μl relevant oligo 3 (10 pmol / ul), 4 μl H2O. ( See table IV, below)

TABLE IVOligo 1 (32P-Experimentlabelled)Oligo 2Oligo 3QAh 1Ah 6Ah 7RAh 5Ah 6Ah 7

[0278] Anneal as described above. Experiment Q is added 1 μl 100 mM EDC and 1 μl 100 mM NHS . Experiment R is added 1 μl 100 mM TSAT. Incubate at 25° C. for about 1.5 h, and then analyze by 10% urea polyacrylamide gel electrophoresis.

[0279] The results are shown in FIG. 4.

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Abstract

The invention relates to a method for synthesizing templated molecules attached to the templated which directed the synthesis thereof. The method involves a template, a scaffold functional entity and a functional entity attached to a building block, which, in turn, is attached the template. The scaffold functional entity and the functional entity of the building block are both provided with complementary dimerization domains allowing the functional entities to come into close proximity when the complementary domains interact with to each other. The method may be used for generating libraries of templated molecules which may be selected for biological activity.

Description

TECHNICAL FIELD OF THE INVENTION [0001] The present invention relates to a method for synthesising templated molecules. The method implies a high local concentration of reactive groups intended to participate in a formation of a linkage, thus increasing the probability of linkage formation. The invention also relates to a library, that is a plurality of templated molecules, wherein each of the. templated molecules are attached to the template which directed the synthesis thereof. BACKGROUND [0002] The generation of molecules carrying new properties remains a challenging task. Recently, a number of procedures have been suggested that should allow a more efficient generation and screening of a larger number of molecules. The approaches taken involve the encoding and / or templating of molecules other than natural biopolymers such as peptide, RNA and DNA. These approaches allow the researcher to generate and screen a huge number of molecules in a short time. This should lead to better mo...

Claims

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Application Information

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IPC IPC(8): F03D9/00C07BC07B61/00C07D405/04C07HC07H21/00C12NC12N15/10C12PC12P1/00C12P19/34C12Q1/68H02P9/00
CPCC07B2200/11C07D405/04C07H21/00C12N15/1068C12Q1/6816C40B40/06C40B40/14C07H21/04
Inventor PEDERSEN, HENRIKHOLTMANN, ANETTEFRANCH, THOMASHAAHR, ALEXFELDING, JAKOB
Owner NUEVOLUTION AS
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